Ready-to-eat salads and berry fruits purchased in Italy contaminated by Cryptosporidium spp., Giardia duodenalis, and Entamoeba histolytica.

Ready-to-eat (RTE) salads and berries are increasingly consumed in industrialized countries. These products can be contaminated by pathogenic parasites that have been responsible for foodborne outbreaks worldwide. In Italy, there are few data on contamination of RTE salads and berries with parasite transmission stages and this requires more-in-depth investigations. To estimate the prevalence of contamination with Cryptosporidium spp. and Giardia duodenalis in these fresh products, a total of 324 packages of local RTE mixed salads - belonging to three different industrial brands - and 324 packages of berries - blueberries from Peru, blackberries from Mexico, raspberries from Italy - were bought from supermarkets located in the Provinces of Bari and Foggia, Apulia, Italy. A pool size of nine packages was chosen and a total of 72 pools were processed in the whole year. After washing, the pellets were examined by microscopy (FLOTAC) and tested using conventional simplex PCR, targeting Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp., and sequencing. Several Cryptosporidium species and Giardia duodenalis assemblages, some of which are of potential zoonotic relevance, as well as Entamoeba spp., were identified in both matrices. By microscopy, Giardia-like cysts in local raspberries and Entamoeba-like cysts in imported blueberries were detected. Giardia duodenalis (Assemblages A, B and E) and Entamoeba histolytica were molecularly confirmed with overall prevalences of 4.6% (95% C.I. 3.0-6.8) and 1% (95% C.I. 0.3-2.1), respectively. Molecular methods identified Cryptosporidium ryanae, Cryptosporidium bovis, Cryptosporidium xiaoi, and Cryptosporidium ubiquitum in both matrices, with a prevalence of 5.1% (95% C.I. 3.3-7.3). A distinct seasonality in prevalence was observed for G. duodenalis, with most positives occurring in spring, whereas Cryptosporidium showed no significant seasonal variations. These results highlight that inadequate management of fresh produce, both locally produced and imported, along the food chain may have the potential for consequences on human health.

Evaluation of the performance of a lateral flow device for quantitative detection of anti-SARS-CoV-2 IgG.

INTRODUCTION: The AbC-19™ lateral flow immunoassay (LFIA) performance was evaluated on plasma samples from a SARS-CoV-2 vaccination cohort, WHO international standards for anti-SARS-CoV-2 IgG (human), individuals ≥2 weeks from infection of RT-PCR confirmed SARS-CoV-2 genetic variants, as well as microorganism serology. METHODS: Pre-vaccination to three weeks post-booster samples were collected from a cohort of 111 patients (including clinically extremely vulnerable patients) from Northern Ireland. All patients received Oxford-AstraZeneca COVID-19 vaccination for the first and second dose, and Pfizer-BioNTech for the third (first booster). WHO international standards, 15 samples from 2 variants of concern (Delta and Omicron) and cross-reactivity with plasma samples from other microorganism infections were also assessed on AbC-19™. RESULTS: All 80 (100%) participants sampled post-booster had high positive IgG responses, compared to 38/95 (40%) participants at 6 months post-first vaccination. WHO standard results correlated with information from corresponding biological data sheets, and antibodies to all genetic variants were detected by LFIA. No cross-reactivity was found with exception of one (of five) Dengue virus samples. CONCLUSION: These findings suggest BNT162b2 booster vaccination enhanced humoral immunity to SARS-CoV-2 from pre-booster levels, and that this antibody response was detectable by the LFIA. In combination with cross-reactivity, standards and genetic variant results would suggest LFIA may be a cost-effective measure to assess SARS-CoV-2 antibody status.

Strengthening of district mental health services in Gauteng Province, South Africa.

In response to the Life Esidimeni tragedy, the Gauteng Department of Health established a task team to advise on the implementation of the Health Ombud's recommendations and to develop a mental health recovery plan. Consistent with international human rights and South African legislation and policy, the plan focused on making mental healthcare more accessible, incorporating a strategy to strengthen district mental health services to deliver community-based care for people with any type and severity of mental illness. The strategy included an organogram with three new human resource teams integrated into the district health system: a district specialist mental health team to develop a public mental health approach, a clinical community psychiatry team for service delivery, and a team to support non-governmental organisation governance. This article discusses the strategy in terms of guiding policies and legislation, the roles and responsibilities of the various teams in the proposed organogram, and its sustainability.

Mental health of healthcare workers during the COVID-19 outbreak: A rapid scoping review to inform provincial guidelines in South Africa.

COVID-19 is prevalent in sub-Saharan Africa. The healthcare response to the pandemic depends upon a mentally and physically healthy workforce. Infectious disease outbreaks cause high psychosocial stress among healthcare workers, which may impact negatively on workplace functioning. To understand which mental health conditions may occur and which interventions could be considered, we conducted a rapid scoping review. Using a 2018 systematic review as the starting point, PubMed, Cochrane, Web of Science and MEDLINE databases were searched for any type of evidence published in English between 2014 and 2020 on mental health of healthcare workers exposed to infectious disease outbreaks; 19 primary studies and 13 opinion pieces were included. Depression, anxiety, post-traumatic stress, and other mental health conditions were noted among healthcare workers exposed to COVID-19 and other outbreaks. Although no effectiveness studies were identified, certain proposed interventions may be implemented by healthcare leaders. Further research is recommended.

Mental health of healthcare workers during the COVID-19 outbreak: A rapid scoping review to inform provincial guidelines in South Africa

COVID-19 is prevalent in sub-Saharan Africa. The healthcare response to the pandemic depends upon a mentally and physically healthy workforce. Infectious disease outbreaks cause high psychosocial stress among healthcare workers, which may impact negatively on workplace functioning. To understand which mental health conditions may occur and which interventions could be considered, we conducted a rapid scoping review. Using a 2018 systematic review as the starting point, PubMed, Cochrane, Web of Science and MEDLINE databases were searched for any type of evidence published in English between 2014 and 2020 on mental health of healthcare workers exposed to infectious disease outbreaks; 19 primary studies and 13 opinion pieces were included. Depression, anxiety, post-traumatic stress, and other mental health conditions were noted among healthcare workers exposed to COVID-19 and other outbreaks. Although no effectiveness studies were identified, certain proposed interventions may be implemented by healthcare leaders. Further research is recommended

An apple a day: an outbreak of cryptosporidiosis in Norway associated with self-pressed apple juice.

In the autumn of 2018, an outbreak of cryptosporidiosis affected adult employees from the same company in Western Norway. The organism was Cryptosporidium parvum, GP60 subtype IIaA14G1R1. All those infected had drunk from the same container of self-pressed apple juice. Incubation period (1 week) and clinical signs were similar among those infected, although some experienced a more prolonged duration of symptoms (up to 2-3 weeks) than others. The infections resulted after consumption from only one of 40 containers of juice and not from any of the other containers. It seems that although Cryptosporidium oocysts were detected in a sample from another container, the contamination did not affect the whole batch. This is perhaps indicative of a restricted contamination event, either from contaminated ground in the orchard, or during collection of the fruit, or during processing. Although outbreaks of food-borne cryptosporidiosis have previously been associated with consumption of contaminated apple juice, most of the more recent outbreaks of food-borne cryptosporidiosis have been associated with salad vegetables or herbs. This outbreak, the first outside USA reported to be associated with apple juice, is a timely reminder that such juice is a suitable transmission vehicle for Cryptosporidium oocysts, and that appropriate hygienic measures are essential in the production of such juice, including artisanal (non-commercial) production.

Apparent absence of Giardia infections among children under 5-years of age with acute watery diarrhoea in Abakaliki, Nigeria.

Although the impact of diarrhoeal disease on paediatric health in Nigeria has decreased in recent years, it remains an important cause of morbidity and mortality in children under 5 years. Rotavirus is recognised as an important aetiological agent, but information on the contribution of intestinal protozoa to watery diarrhoea in this age group in Nigeria is scarce. In this cross-sectional study, faecal samples from children admitted to healthcare centres in Abakaliki, Nigeria with acute watery diarrhoea (N = 199) and faecal samples from age-matched controls (N = 37) were examined for Cryptosporidium and Giardia using immunofluorescent antibody testing and molecular methods. Cryptosporidium was identified in 13 case samples (6.5%) and no control samples. For three samples, molecular characterisation indicated C. hominis, GP60 subtypes IaA30R3, IaA14R3 and IdA11. Giardia was not detected in any samples. This contrast in prevalence between the two intestinal protozoa may reflect their variable epidemiologies and probably differing routes of infection. Given that these two parasitic infections are often bracketed together, it is key to realise that they not only have differing clinical spectra but also that the importance of each parasite is not the same in different age groups and/or settings.

Mortality analysis of people with severe mental illness transferred from long-stay hospital to alternative care in the Life Esidimeni tragedy.

A mortality analysis of the Life Esidimeni tragedy was precluded during the investigation by the Health Ombud by the lack of data integrity. Information on the mental healthcare users (MHCUs) transferred out of Life Esidimeni hospitals between October 2015 and June 2016 was subsequently collected by the Gauteng Department of Health, permitting statistical analysis. Survival rates were calculated according to gender and transfer destination and adjusted for patient age. Mortality was compared with that of the general population for the calendar year of 2016. Of the 1 442 MHCUs, 15% were transferred to specialised psychiatric hospitals and 85% to a rehabilitation centre or non-governmental residential facility. By the end of August 2017, 9% (n=131) of the cohort had died. Significant predictors of survival were younger age (p<0.0001) and transfer to a psychiatric institution (p=0.004). The age-adjusted death rate was 63/1 000 and the overall standardised mortality ratio (SMR) was 4.9 (95% confidence interval (CI) 3.92 - 5.80), with an SMR of 3.9 (95% CI 2.95 - 4.86) for men and 6.3 (95% CI 4.22 - 8.38) for women. The excess deaths are therefore quantified, and the high-risk environment of the rehabilitation centre and residential facilities confirmed. High mortality among MHCUs is unlikely to be confined to the Life Esidimeni tragedy; monitoring of preventable deaths in this vulnerable population is recommended.

Giardia duodenalis in primates: Classification and host specificity based on phylogenetic analysis of sequence data.

Giardia duodenalis colonizes the gastrointestinal tract of a wide range of hosts, including humans and other primates. It is grouped into eight different Assemblages and, beyond that, into a number of sub-Assemblages, defined ad hoc on the basis of genetic differences; these various groups are often considered to be associated with a specific restricted host range. The aim of this study was to use publicly available genotyping data to investigate the relatedness of human and non-human primate (NHP) Giardia isolates in order to evaluate the usefulness of current taxonomic classification and to assess whether there is potential for zoonotic transmission between humans and NHP. Our final data set consisted of sequence data from 165 isolates, 111 from NHP and 54 from humans. Assemblages were well defined, but sub-Assemblages across Assemblage B were not resolved. Although sub-Assemblages AI and AII were resolved, the terms were not found to capture any useful molecular or host/deme properties. In the phylogenetic tree, NHP isolates were scattered among human isolates across Assemblages A and B, and were even found in Assemblage E. We conclude that there does not appear to be significant molecular distinction between human and NHP Giardia isolates across these four molecular markers. Thus, on the basis of these markers, we cannot exclude a risk for zoonotic and anthropozoonotic transmission of Assemblages A and B isolates, irrespective of sub-Assemblage classification. We further evaluated the relative merit of the four genes used in genotyping studies. The tpi, gdh and bg genes gave relatively congruent tree topologies, but the SSU gene did not resolve Assemblages according to the current classification. Future genotyping efforts should aim for multilocus or whole-genome approaches and, in particular, use of the SSU gene as the sole marker should be avoided when possible.

Changes in the seroprevalence of cysticercosis in suspected patients in Chandigarh, India between 1998 and 2014: analysis of 17 years of data.

Changes in seroprevalence of cysticercosis diagnosed in Chandigarh, India between 1998 and 2014 were investigated by extraction and analysis of data from records held at the Postgraduate Institute of Medical Education and Research in Chandigarh. Among the total number of samples for which cysticercosis had been suspected during this period (N = 9650), 1716 (17·8%) were seropositive. Adults were more likely to be seropositive than children, and women were more likely to be seropositive than men. In addition to there being fewer patients with suspicion of cysticercosis over the data analysis period, the proportion of patients seropositive also reduced significantly. Despite these reductions, which are probably associated with improved infrastructure and sanitation within Chandigarh, and despite meat consumption being relatively rare in this area, the extent of cysticercosis in this population remains problematic. Further efforts should be made to reduce transmission of this infection, with particular emphasis on women. Such efforts should follow the One Health concept, and involve medical efforts (including diagnosis and treatment of T. solium tapeworm carriers), veterinary efforts directed towards meat inspection and prevention of infection of pigs, and environmental health and sanitation engineers (to minimize environmental contamination with human waste).

Symptomatic and asymptomatic secondary transmission of Cryptosporidium parvum following two related outbreaks in schoolchildren.

Two related outbreaks (in 2009 and 2012) of cryptosporidiosis in Norwegian schoolchildren during a stay at a remote holiday farm provided us with a natural experiment to investigate possible secondary transmission of Cryptosporidium parvum IIa A19G1R1. After the children had returned home, clinical data and stool samples were obtained from their household contacts. Samples were investigated for the presence of Cryptosporidium oocysts by immunofluorescence antibody test. We found both asymptomatic and symptomatic infections, which are likely to have been secondary transmission. Laboratory-confirmed transmission rate was 17% [4/23, 95% confidence interval (CI) 7·0-37·1] in the 2009 outbreak, and 0% (95% CI 0-16·8) in the 2012 outbreak. Using a clinical definition, the probable secondary transmission rate in the 2012 outbreak was 8% (7/83, 95% CI 4·1-16·4). These findings highlight the importance of hygienic and public health measures during outbreaks or individual cases of cryptosporidiosis. We discuss our findings in light of previous studies reporting varying secondary transmission rates of Cryptosporidium spp.

Molecular identification of Neospora caninum from calf/foetal brain tissue and among oocysts recovered from faeces of naturally infected dogs in southern Ethiopia.

This study sought to confirm and investigate further recently published information regarding the occurrence of Neospora caninum in cattle in Ethiopia and investigate infection in dogs, the canine definitive host, in this region. Faecal samples from 383 dogs in Hawassa, Ethiopia were examined by microscopy for Neospora-like oocysts, and positive samples then analysed by a molecular approach (DNA isolation, PCR and sequencing at the ITS1 gene). Brain tissue samples from four late term aborted foetuses, one congenitally defective calf (hind leg arthrogryposis) and placental tissue from cattle in the same area were also examined by the same molecular approach. All foetal, calf and placental tissue were associated with Neospora seropositive dams. A high prevalence of Neospora-like oocysts (11.5 µm±1.5 µm diameter) was observed in faecal samples from dogs (37 positive samples; 9.7% prevalence), and in 17 of these the identification was confirmed by PCR, giving a prevalence of confirmed infection of 4.4%. N. caninum DNA was also detected in all foetal and calf brain tissue samples. Sequencing revealed only minor differences among all PCR products, whether from oocysts or from brain tissue samples. These data provide molecular evidence of the presence of N. caninum infection in both dog and cattle in this region of Ethiopia. Moreover these findings highlight the role of dogs in maintaining and spreading the infection horizontally in the study area. The high frequency of N. caninum infection in household dogs as well as farm dogs is worthy of further investigation.

Second outbreak of infection with a rare Cryptosporidium parvum genotype in schoolchildren associated with contact with lambs/goat kids at a holiday farm in Norway.

In March 2012, a second outbreak of Cryptosporidium parvum affected children following a stay at a holiday farm in Norway; the first outbreak occurred in 2009. We studied a cohort of 145 schoolchildren who had visited the farm, of which 40 (28%) were cases. Cryptosporidium oocysts were detected in faecal samples from humans, goat kids and lambs. Molecular studies revealed C. parvum subtype IIa A19G1R1 in all samples including human samples from the 2009 outbreak. A dose-response relationship was found between the number of optional sessions with animals and illness, increasing from two sessions [risk ratio (RR) 2·7, 95% confidence interval (CI) 0·6-11·5] to six sessions (RR 8·0, 95% CI 1·7-37·7). The occurrence of two outbreaks 3 years apart, with the same subtype of C. parvum, suggests that the parasite is established in the farm's environment. We recommend greater emphasis on hand hygiene and routines related to animal contact.

Reproductive disorders in relation to Neospora caninum, Brucella spp. and bovine viral diarrhoea virus serostatus in breeding and dairy farms of central and southern Ethiopia.

Abortion and stillbirth are important reproductive disorders in the dairy industry and are often caused by infectious agents. This study investigated whether bovine viral diarrhoea virus (BVDV), Brucella spp., and Neospora caninum are associated with abortion and/or stillbirth in dairy cattle in Ethiopia. Dairy cattle from 99 farms were categorized as cases (n=134) or controls (n=268) according to reproductive data. Blood samples were screened for antibodies for these infectious agents. The overall proportion of cattle that were seropositive for BVDV, Brucella spp., and N. caninum was 11∙7%, 3∙2%, and 17∙2%, respectively. Seropositivity for BVDV and Brucella spp. was similar for cases and controls, but significantly more cases were seropositive for N. caninum (29∙8%) than controls (10∙8%). This is the first report demonstrating N. caninum is common in dairy cattle in Ethiopia, and is probably a greater impediment to reproductive success in Ethiopian dairy farms than either BVDV or Brucella spp.

Seroprevalence of Neospora caninum and associated risk factors in intensive or semi-intensively managed dairy and breeding cattle of Ethiopia.

A cross-sectional study of Neospora caninum infection was conducted in major milksheds of Ethiopia. Cattle (n=2334) from 273 farms were bled and the sera screened for antibodies against N. caninum using a commercial ELISA kit. Herd and individual animal level data were collected from farm records and a semi-structured questionnaire format. The overall animal level seroprevalence was 13.3%, while the prevalence at farm level was 39.6%. In urban and peri-urban smallholder dairy farms, the seroprevalence was 14.9%, while 12.9% and 9.8% reactors were found among commercial dairy farms and breeding cattle, respectively. At farm level, 35.7% of urban and peri-urban farms, 47.5% of the commercial farms and five of the breeding farms were found to have at least one infected animal. Purchased cows (OR: 2.3) and cows with history of maternal reproductive disorders (OR: 2.1) were associated with seropositivity at the individual animal level. Crossbred cattle (Holstein-Friesian crossed with indigenous zebu) were associated with lower risk than pure breeds (OR: 0.6). A trend of prevalence increment was observed for large herd sizes (OR: 1.8). Other factors that were associated with seropositivity were: presence of farm dogs for more than 5 years (OR: 1.9), access to farm by wild carnivores (OR: 3.1) and compromised farm hygienic status (OR: 3.4). Abortion, retention of foetal membrane and metritis were the most frequently reported clinical reproductive disorders among seropositive cattle. Together, these finding indicate that N. caninum infection is highly prevalent, widely distributed and clinically important in dairy and breeding cattle of Ethiopia. N. caninum should be considered an important infectious cause of reproductive disorders in Ethiopian cattle, and the risk factors for exposure identified here should be used as basis for implementing control measures that could limit the transmission of this infection.

European pet travel: misleading information from veterinarians and government agencies.

Inter-country travel of companion animals provides an opportunity for introduction of zoonotic pathogens, such as rabies virus and Echinococcus spp. Regulations are in place to control this threat, but Schengen Agreements mean that border controls between some European countries are minimal, and animals may enter countries without any checks that they have been appropriately treated. Veterinarians provide an important source of information for people intending to travel with their pets. We conducted a telephone survey to investigate provision of correct advice to someone intending to travel with their dog to Norway. Mainland Norway is considered free of both rabies and E. multilocularis and is a signatory to the Schengen Agreement. Ten randomly selected veterinary clinics were surveyed in Austria, Belgium (Wallonia), Finland, France, Germany, Norway, Sweden, Switzerland and United Kingdom. The information provided was scored as correct, incorrect or incomplete. The information provided by secondary information sources (website or government agency), which the clinic had referred the caller to, was also assessed (correct, incorrect, incomplete). Whilst the majority of clinics provided appropriate information regarding rabies, many clinics did not provide correct information regarding treatment for E. multilocularis. Less than one in 10 clinics provided the correct information regarding both pathogens directly at the time of calling. The correct information was obtained, once taking into account secondary sources, just 62% of the time. Countrywise, most clinics in Finland provided correct advice, either directly or indirectly via referring the caller to another source, whilst the majority in Belgium, Germany and France did not. The apparent paucity of readily accessible, correct advice for owners intending to travel with their dogs is concerning. The compulsory treatment regulations are only as good as the checks that ensure compliance, and this is also lacking in some countries.

Unexpected results from large-scale cryptosporidiosis screening study in calves in Tanzania.

A study was undertaken to investigate Cryptosporidium infection in crossbreed dairy calves in two districts in Tanzania. A total of 943 fecal samples from 601 dairy calves were included in the study, with calves from both smallholder dairy farms and from large-scale and medium-scale dairy farms. The modified Ziehl-Neelsen (mZN) technique was used to examine 710 samples, and 13 of these were considered to be positive for Cryptosporidium. These 13 samples considered positive by mZN, along with the remaining 233 samples, were analysed by immunofluorescent antibody test (IFAT). Of these 246 samples examined by IFAT, 15 samples, 10 of which were considered positive by mZN, were also examined by the auramine phenol technique, and 5 samples, all of which were considered positive by mZN, were analysed by PCR. The results from the IFAT, auramine phenol and PCR analyses demonstrated that none of the samples contained Cryptosporidium oocysts, indicating that, cryptosporidiosis is currently not a problem in dairy calves in these regions of Tanzania. These unexpected results are discussed with respect to other reports on cryptosporidiosis in calves that suggest that this parasite is a serious calf disease globally, and particularly in relation to studies from Tanzania. We suggest that results from studies of cattle in Tanzania, in which mZN has been used as the sole analytical method, should be treated with caution.

The zoonotic potential of Giardia and Cryptosporidium in Norwegian sheep: a longitudinal investigation of 6 flocks of lambs.

Faecal samples collected from lambs on 6 Norwegian farms on 2 separate occasions during spring/summer 2008 (approximately 550 samples collected at each occasion) were examined for Cryptosporidium oocysts and Giardia cysts by immunofluorescent antibody test. Overall prevalence at the first sampling was approximately 23% for Giardia and 15% for Cryptosporidium, and at the second sampling approximately 31% for Giardia and 24% for Cryptosporidium, with substantial between-farm variation on each sampling occasion. To assess the potential public health significance of these infections, molecular analyses were conducted on 42 Giardia isolates and 42 Cryptosporidium isolates, with PCR targeted at one or both of two genes (Giardia: glutamate dehydrogenase and beta-giardin genes; Cryptosporidium: SSU rRNA and actin genes) for each parasite. Of the Giardia isolates, 41 were Assemblage E (non-zoonotic) and 1 was Assemblage B (zoonotic). Of the Cryptosporidium isolates, 35 were cervine genotype (potentially zoonotic) and 7 C. xiaoi (non-zoonotic). These results suggest that sheep in Norway are unlikely to be an important reservoir of zoonotic Giardia in Norway, but might have some public health significance with respect to Cryptosporidium.